Native bromoperoxidases do not bind to nitrocellulose: use of DEAE-cellulose as an alternative in blotting.
Bromoperoxidases were investigated by protein blotting after polyacrylamide gel electrophoresis. While the denatured proteins bound to nitrocellulose, the native enzymes did not. Instead, they could be transferred successfully to DEAE-cellulose. Procedures for immunostaining and glycoprotein detection with concanavalin A on DEAE-cellulose are described. The results indicate that binding of native proteins to nitrocellulose can not necessarily be assumed. DEAE-cellulose is suitable both to investigate this phenomenon or as a substitute for nitrocellulose in blotting of native proteins.
Jordan
P
P
Institut für Pharmazeutische Biologie, Universität Bonn, Federal Republic of Germany.
Vilter
H
H
D015153:Blotting, Western; D003101:Collodion; D003208:Concanavalin A; D003636:DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis; D010544:Peroxidases / Q000032:analysis*; D000459:Phaeophyceae / Q000201:enzymology
D006023:Glycoproteins; D003208:Concanavalin A; D003101:Collodion; D003636:DEAE-Cellulose; D010544:Peroxidases; C026195:bromide peroxidase